Heterologous abs from llama and chicken egg yolk to prevent rotavirus diarrhea

描述（由申请方提供）：我们将探索口服不同的被动异源抗体（Ab）、鸡蛋黄IgY和源自美洲驼重链Ab的重组VHH片段作为轮状病毒（RV）腹泻预防和治疗疫苗接种的替代或补充方法。将在RV疾病的两种新生动物模型（gnotobiotic（Gn）猪和小牛）中评价被动治疗。该研究将在美国俄亥俄州州立大学Saif博士实验室（Gn猪）和阿根廷INTA与Parreqo博士（新生小牛）合作同时进行，作为NIAID的扩展，NIH批准号R 01 AI 033561 -11（6/25/03-12/31/08）。Saif博士与Parreqo博士和费尔南德斯博士（INTA）之间的持续和长期合作促进了最先进的技术转让，用于诊断（检测，试剂）和治疗（疫苗）牛RV感染，这是阿根廷的主要农产品。通过该提案，我们继续在两种易患RV腹泻的新生动物中探索新的免疫学概念（被动异源Ab对RV主动Ab反应的影响）和新的治疗方法（被动IgY，VHH），从而改善反刍动物和人类健康相关的合作：具有正常肠道微生物植物群的常规小牛和缺乏微生物植物群的Gn猪。国外合作者将生产抗体（美洲驼VHH、鸡蛋IgY、牛IgG）并进行小牛实验。来自INTA的学生将在Saif博士的实验室接受培训，学习更新的免疫学程序（流式细胞术等），并与专门的动物疾病模型（Gn猪）合作，以及提高与粘膜免疫相关的分析技能。因此，INTA团队将接受与Gn猪粘膜免疫反应研究相关的试剂和培训，这些试剂和培训将应用于小牛实验。我们的合作也将进一步提高INTA作为拟议领域基础和转化研究卓越中心的声誉。开发来自美洲驼的重组Ab将使得这种天然动物能够可持续地应用于开发用于治疗这种疾病和其他疾病的VHH Ab。这将有利于美洲驼畜牧业生产，以及阿根廷或南美洲其他从事骆驼生产的较贫穷发展中国家（秘鲁、玻利维亚等）的区域微观经济。这些异源被动Ab治疗作为早产儿或严重RV爆发环境（医院，日托中心，发展中国家的农村地区）的补充剂将产生深远的影响。它们代表了在RV疫苗使用不可能、太昂贵、太迟的情况下减少RV腹泻严重程度和死亡的补充治疗;或治疗婴儿和反刍动物疫苗失败的病例。公共卫生相关性：轮状病毒（RV）是全世界幼龄动物和人类婴儿腹泻的主要原因。目前，动物中的RV预防是基于灭活RV母体疫苗以增强被动免疫，而减毒活RV疫苗最近已被批准用于人类婴儿。疫苗的有效性和安全性仍然不清楚，特别是在发展中国家。我们将探索口服不同的被动异源抗体，骆驼重链抗体片段（VHH）和鸡蛋黄IgY作为替代/互补的方法来控制RV腹泻的动物和人类，在两个动物模型的轮状病毒感染和疾病gnotobiotic（Gn）的猪和新生小牛。如果我们的异源被动抗体（Ab）策略有效，这些治疗将对早产儿或严重RV爆发环境的补充产生最深远的影响（医院、日托中心、发展中国家的农村地区），提供一种替代的快速治疗方法，以减少RV腹泻的严重程度和在RV疫苗使用不可能、太昂贵、太晚的情况下的死亡;或治疗疫苗失效的病例。

DESCRIPTION (provided by applicant): We will explore oral administration of different passive heterologous antibodies (Abs), chicken egg yolk IgY and recombinant VHH fragments derived from llama heavy chain Abs as an alternative or complementary approach to vaccination for rotavirus (RV) diarrhea prevention and treatment. The passive treatments will be evaluated in two neonatal animal models of RV disease, gnotobiotic (Gn) pigs and calves. The research will be done simultaneously, in the US at Dr. Saif's Lab, The Ohio State University (Gn pig) and in Argentina, at INTA in collaboration with Dr. Parreqo (neonatal calves), as an extension of NIAID, NIH Grant No. R01 AI033561-11 (6/25/03-12/31/08). The ongoing and long-term collaboration between Dr. Saif and Drs. Parreqo and Fernandez (INTA) has facilitated transfer of state-of-the-art technology for diagnosis (assays, reagents) and treatments (vaccines) for RV infections of cattle, the major agricultural commodity in Argentina. Through this proposal we continue this collaboration related to improving both ruminant and human health by exploring new immunologic concepts (impact of passive heterologous Ab on active Ab responses to RV) and new treatments (passive IgY,VHH) in two neonatal animals susceptible to RV diarrhea: conventional calves with a normal gut microbial flora and Gn pigs lacking the microbial flora. The foreign collaborator will produce the Abs (llama VHH, chicken egg IgY, bovine IgG) and conduct the calf experiments. Students from INTA will be trained in Dr. Saif's lab to learn newer immunologic procedures (flow cytometry, etc) and work with specialized animal disease models (Gn pigs) as well as to enhance analytical skills related to mucosal immunity. Thus the INTA team will receive reagents and training related to the study of mucosal immune responses in Gn pigs that will be applied to the calf experiments. Our collaboration will also further INTA's reputation as a Center of Excellence for basic and translational studies in the proposed areas. Development of recombinant Abs from llamas, will enable a sustainable application of this native animal for the development of VHH Ab applied to treat this and other diseases. This will benefit llama livestock production, as well as regional micro-economies for Argentina or other poorer developing countries of South America (Peru, Bolivia, etc) engaged in Camelid production. These heterologous passive Ab treatments would have profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries). They represent complementary treatments to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too expensive, too late; or to treat cases of vaccine failure for both infants and ruminants. PUBLIC HEALTH RELEVANCE: Rotavirus (RV) is the leading cause of diarrhea in young animals and human infants, worldwide. Currently, RV prevention in animals is based on inactivated RV maternal vaccines to enhance passive immunity, whereas live-attenuated RV vaccines have been recently approved for human infants. Vaccine efficacy and safety remain unclear especially in developing countries. We will explore oral administration of different passive heterologous antibodies, llama heavy chain antibody fragments (VHH) and chicken egg yolk IgY as alternative/complementary approaches to control RV diarrhea for both animals and humans, in two animal models of rotavirus infection and disease gnotobiotic (Gn) pigs and neonatal calves. If our heterologous passive antibody (Ab) strategies are effective, these treatments would have the most profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries), by providing an alternative rapid treatment to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too costly, too late; or to treat cases of vaccine failure.